Differentiation of human pluripotent stem cells (hPSCs) requires precise signaling control at each step to mimic the embryonic development. Advances in using highly specific and potent small molecule modulators of signaling pathways have allowed efficient generation of nearly homogenous populations of specific cell types from hPSCs. For highly efficient conversion of hESCs/iPSCs into neural precursor cells in a monolayer fashion, inhibition of both BMP and TGFβ signaling pathways (i.e., dual SMAD inhibition) by small molecule inhibitors, LDN-193189 (0.1 µM) and SB431542 (10 µM) respectively (i.e., Neural-LSB), was developed as the most effective condition. Under this dual SMAD inhibition using the Neural-LSB small molecule cocktail, more than 80% of hPSCs are rapidly induced into Pax6+ neural precursor cells. This condition can be further combined with other patterning cues to generate subtype specific neural and neuronal cells, such as floor plate and midbrain dopaminergic neurons using Hedgehog and Wnt pathway activators (i.e., purmorphamine and CHIR99021), or nociceptors using 3i (combination of CHIR99021/GSK3 inhibitor, SU5402/FGF-VEGF-PDGF inhibitor, DAPT/Notch inhibitor). 

XcessBio’s Neural-LSB contains the combination of LDN193189/BMP inhibitor and SB431542/TGFβ inhibitor.

How to Use:

• In vitro: Simply add/dilute the received LSB stock solutions at 1:1000 into your hESC/iPSC differentiation basal media to make the LSB induction media and use it to initiate neural differentiation.

Reference:

• 1.  Chambers, S.M. et al. Combined small-molecule inhibition accelerates developmental timing and converts human pluripotent stem cells into nociceptors. (2012) Nature Biotechnology 30, 715-720.

• 2.  Chambers, S.M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. (2009) Nat. Biotechnol. 27, 275–280.

• 3.  Yu, P.B. et al. BMP type I receptor inhibition reduces heterotopic [corrected] ossification. (2008) Nat. Med. 14, 1363–1369.

Products are for research use only. Not for human use.